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. Author manuscript; available in PMC: 2012 Aug 15.
Published in final edited form as: J Immunol. 2011 Jul 6;187(4):1797–1806. doi: 10.4049/jimmunol.1002628

Figure 1. Effect of RANKL and IL-4 on the rapid activation of NF-κB.

Figure 1

Primary BMM were cultured in the presence or absence of RANKL (150 ng/ml) or IL-4 (10 ng/ml) as indicated for 30 minutes. Cytoplasmic (C) and nuclear (N) lysates were prepared as described in the Methods section. A. Lysates were analyzed by western blotting with anti-IκBα or anti-p50 as indicated. B. Top panel. Nuclear lysates were incubated with the biotin-labeled κB oligo and analyzed by EMSA assay as described in the Methods section. Total lysates of 293T cells transfected with p50 and p65 were run simultaneously as reference. Bottom panel, Nuclear lysates were analyzed by western blotting using anti-p65, anti-p50, anti-STAT6, or anti-lamin A antibodies. C. BMM were pretreated with 10 ng/ml IL-4 for various times and then were treated with RANKL (150 ng/ml) or IL-4 (10 ng/ml) as indicated for 30 minutes. Nuclear lysates were harvested and analyzed using anti-p50, anti-p65, or anti-lamin A antibodies.